rtest  (Solis BioDyne)


Bioz Verified Symbol Solis BioDyne is a verified supplier
Bioz Manufacturer Symbol Solis BioDyne manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95

    Structured Review

    Solis BioDyne rtest
    Optimization, analytical sensitivity and clinical performance of <t>rTEST</t> COVID‐19/FLU qPCR kit. A. Heatmaps illustrate combinatorial testing of two IAV primer/probe sets at either high or low viral input (10 000 versus 10 copies per reaction). B. Heatmaps show combinatorial testing of IBV primer/probe sets at low viral input (10 copies per reaction). In panels A, B, the best performing primer/probe combinations (highlighted by green rectangles) were selected based on C t (darker colours denote higher sensitivity), fluorescent intensity (∆ R , lighter colours correspond to higher intensity) and the number of replicates that amplified. C. Analytical sensitivity of the <t>multiplexed</t> <t>SARS‐CoV‐2</t> E and RdRP (both labelled with FAM), IAV PB1 and IBV PA (both labelled with YY), and RNase P assay. The dotted line at C t = 40 serves as a threshold after which amplification is considered invalid. D. Assessment of competitive interference of 1000 copies of IAV per reaction (500× LoD) on the analytical sensitivity of the SARS‐CoV‐2 E and RdRP assays multiplexed together. E. Assessment of competitive interference of 1000 copies of SARS‐CoV‐2 per reaction (500× LoD) on the analytical sensitivity of the IAV PB1 assay. F. Clinical performance of the rTEST COVID‐19/FLU qPCR kit. The dotted line and shaded area indicate samples that were not detected by a particular assay. C t , cycle threshold; E, envelope gene; IAV, influenza A; IBV, influenza B; PA, polymerase acidic protein; PB1, polymerase basic 1 protein; ND, not detected within 45 cycles; NTC, no template control; RdRP, RNA‐dependent RNA polymerase; Δ R , normalized fluorescent intensity.
    Rtest, supplied by Solis BioDyne, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rtest/product/Solis BioDyne
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rtest - by Bioz Stars, 2022-08
    95/100 stars

    Images

    1) Product Images from "Sequential development of several RT‐qPCR tests using LNA nucleotides and dual probe technology to differentiate SARS‐CoV‐2 from influenza A and B"

    Article Title: Sequential development of several RT‐qPCR tests using LNA nucleotides and dual probe technology to differentiate SARS‐CoV‐2 from influenza A and B

    Journal: Microbial Biotechnology

    doi: 10.1111/1751-7915.14031

    Optimization, analytical sensitivity and clinical performance of rTEST COVID‐19/FLU qPCR kit. A. Heatmaps illustrate combinatorial testing of two IAV primer/probe sets at either high or low viral input (10 000 versus 10 copies per reaction). B. Heatmaps show combinatorial testing of IBV primer/probe sets at low viral input (10 copies per reaction). In panels A, B, the best performing primer/probe combinations (highlighted by green rectangles) were selected based on C t (darker colours denote higher sensitivity), fluorescent intensity (∆ R , lighter colours correspond to higher intensity) and the number of replicates that amplified. C. Analytical sensitivity of the multiplexed SARS‐CoV‐2 E and RdRP (both labelled with FAM), IAV PB1 and IBV PA (both labelled with YY), and RNase P assay. The dotted line at C t = 40 serves as a threshold after which amplification is considered invalid. D. Assessment of competitive interference of 1000 copies of IAV per reaction (500× LoD) on the analytical sensitivity of the SARS‐CoV‐2 E and RdRP assays multiplexed together. E. Assessment of competitive interference of 1000 copies of SARS‐CoV‐2 per reaction (500× LoD) on the analytical sensitivity of the IAV PB1 assay. F. Clinical performance of the rTEST COVID‐19/FLU qPCR kit. The dotted line and shaded area indicate samples that were not detected by a particular assay. C t , cycle threshold; E, envelope gene; IAV, influenza A; IBV, influenza B; PA, polymerase acidic protein; PB1, polymerase basic 1 protein; ND, not detected within 45 cycles; NTC, no template control; RdRP, RNA‐dependent RNA polymerase; Δ R , normalized fluorescent intensity.
    Figure Legend Snippet: Optimization, analytical sensitivity and clinical performance of rTEST COVID‐19/FLU qPCR kit. A. Heatmaps illustrate combinatorial testing of two IAV primer/probe sets at either high or low viral input (10 000 versus 10 copies per reaction). B. Heatmaps show combinatorial testing of IBV primer/probe sets at low viral input (10 copies per reaction). In panels A, B, the best performing primer/probe combinations (highlighted by green rectangles) were selected based on C t (darker colours denote higher sensitivity), fluorescent intensity (∆ R , lighter colours correspond to higher intensity) and the number of replicates that amplified. C. Analytical sensitivity of the multiplexed SARS‐CoV‐2 E and RdRP (both labelled with FAM), IAV PB1 and IBV PA (both labelled with YY), and RNase P assay. The dotted line at C t = 40 serves as a threshold after which amplification is considered invalid. D. Assessment of competitive interference of 1000 copies of IAV per reaction (500× LoD) on the analytical sensitivity of the SARS‐CoV‐2 E and RdRP assays multiplexed together. E. Assessment of competitive interference of 1000 copies of SARS‐CoV‐2 per reaction (500× LoD) on the analytical sensitivity of the IAV PB1 assay. F. Clinical performance of the rTEST COVID‐19/FLU qPCR kit. The dotted line and shaded area indicate samples that were not detected by a particular assay. C t , cycle threshold; E, envelope gene; IAV, influenza A; IBV, influenza B; PA, polymerase acidic protein; PB1, polymerase basic 1 protein; ND, not detected within 45 cycles; NTC, no template control; RdRP, RNA‐dependent RNA polymerase; Δ R , normalized fluorescent intensity.

    Techniques Used: Real-time Polymerase Chain Reaction, Amplification

    Schematic illustrating SARS‐CoV‐2 genome and regions targeted by RT‐qPCR primers and probes. A. Schematic overview portrays the SARS‐CoV‐2 genome with RdRP and E gene regions magnified to show the locations of primers and probes of the original Charité protocol, vDetect (v1 and v2), and rTEST RT‐qPCR assays. F, forward primer; P, probe; R, reverse primer. The inset boxes (from left to right) illustrate a SARS‐CoV‐2 particle with labelled structural proteins and RNA, legend describing panel A and the primers and probes used in each test to detect RNase P subunit p30 (RPP30). B. Diagram compares the sequences of RdRP and E gene primers and probes for the original Charité protocol, vDetect (v.1) and vDetect v.2 and rTEST RT‐qPCR assays to the Wuhan reference sequence. The numbers written above the Wuhan reference sequence correspond to the start and end base positions of the sequence Reverse primer sequences are written in the reverse complement (rc). Magenta lines and letters represent mixed bases found in the primers and probes in the Charité protocol that were replaced with the correct bases in vDetect v1 (blue lines and letters). Red lines and letters signify LNA‐modified bases.
    Figure Legend Snippet: Schematic illustrating SARS‐CoV‐2 genome and regions targeted by RT‐qPCR primers and probes. A. Schematic overview portrays the SARS‐CoV‐2 genome with RdRP and E gene regions magnified to show the locations of primers and probes of the original Charité protocol, vDetect (v1 and v2), and rTEST RT‐qPCR assays. F, forward primer; P, probe; R, reverse primer. The inset boxes (from left to right) illustrate a SARS‐CoV‐2 particle with labelled structural proteins and RNA, legend describing panel A and the primers and probes used in each test to detect RNase P subunit p30 (RPP30). B. Diagram compares the sequences of RdRP and E gene primers and probes for the original Charité protocol, vDetect (v.1) and vDetect v.2 and rTEST RT‐qPCR assays to the Wuhan reference sequence. The numbers written above the Wuhan reference sequence correspond to the start and end base positions of the sequence Reverse primer sequences are written in the reverse complement (rc). Magenta lines and letters represent mixed bases found in the primers and probes in the Charité protocol that were replaced with the correct bases in vDetect v1 (blue lines and letters). Red lines and letters signify LNA‐modified bases.

    Techniques Used: Quantitative RT-PCR, Sequencing, Modification

    Optimization of room temperature stable kit and dual probes for rTEST COVID‐19 qPCR kit. (A, B) Graphs depict the effects of decoy nucleic acids (tRNA or salmon sperm DNA) or pure oligonucleotides (pure) on the stability of lyophilized positive control left at room temperature over a one‐month period as assessed by amplification of RdRP (A) and E (B) genes. (C) Performance of either fresh or an rTEST COVID‐19 qPCR kit left a t room temperature for 1‐month on amplification of SARS‐CoV‐2 E and RdRP genes and human RNase P. Comparison of analytical sensitivity (D, F) and fluorescent intensity (E, G) between single probes versus dual probes for both RdRP (D, E) and E (F, G) genes. Dotted line at C t = 40 serves as a threshold after which amplification is considered invalid. * P
    Figure Legend Snippet: Optimization of room temperature stable kit and dual probes for rTEST COVID‐19 qPCR kit. (A, B) Graphs depict the effects of decoy nucleic acids (tRNA or salmon sperm DNA) or pure oligonucleotides (pure) on the stability of lyophilized positive control left at room temperature over a one‐month period as assessed by amplification of RdRP (A) and E (B) genes. (C) Performance of either fresh or an rTEST COVID‐19 qPCR kit left a t room temperature for 1‐month on amplification of SARS‐CoV‐2 E and RdRP genes and human RNase P. Comparison of analytical sensitivity (D, F) and fluorescent intensity (E, G) between single probes versus dual probes for both RdRP (D, E) and E (F, G) genes. Dotted line at C t = 40 serves as a threshold after which amplification is considered invalid. * P

    Techniques Used: Real-time Polymerase Chain Reaction, Positive Control, Amplification

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Solis BioDyne soliscript 1 step cov kit
    Optimization, analytical sensitivity and clinical performance of rTEST COVID‐19/FLU qPCR <t>kit.</t> A. Heatmaps illustrate combinatorial testing of two IAV primer/probe sets at either high or low viral input (10 000 versus 10 copies per reaction). B. Heatmaps show combinatorial testing of IBV primer/probe sets at low viral input (10 copies per reaction). In panels A, B, the best performing primer/probe combinations (highlighted by green rectangles) were selected based on C t (darker colours denote higher sensitivity), fluorescent intensity (∆ R , lighter colours correspond to higher intensity) and the number of replicates that amplified. C. Analytical sensitivity of the multiplexed <t>SARS‐CoV‐2</t> E and RdRP (both labelled with FAM), IAV PB1 and IBV PA (both labelled with YY), and RNase P assay. The dotted line at C t = 40 serves as a threshold after which amplification is considered invalid. D. Assessment of competitive interference of 1000 copies of IAV per reaction (500× LoD) on the analytical sensitivity of the SARS‐CoV‐2 E and RdRP assays multiplexed together. E. Assessment of competitive interference of 1000 copies of SARS‐CoV‐2 per reaction (500× LoD) on the analytical sensitivity of the IAV PB1 assay. F. Clinical performance of the rTEST COVID‐19/FLU qPCR kit. The dotted line and shaded area indicate samples that were not detected by a particular assay. C t , cycle threshold; E, envelope gene; IAV, influenza A; IBV, influenza B; PA, polymerase acidic protein; PB1, polymerase basic 1 protein; ND, not detected within 45 cycles; NTC, no template control; RdRP, RNA‐dependent RNA polymerase; Δ R , normalized fluorescent intensity.
    Soliscript 1 Step Cov Kit, supplied by Solis BioDyne, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/soliscript 1 step cov kit/product/Solis BioDyne
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    soliscript 1 step cov kit - by Bioz Stars, 2022-08
    95/100 stars
      Buy from Supplier

    94
    Solis BioDyne 1 step multiplex probe kit
    Principle of the RKI/ZBS1 protocol for the real-time PCR detection of SARS-CoV-2. RKI/ZBS1 SARS-CoV-2 protocol
    1 Step Multiplex Probe Kit, supplied by Solis BioDyne, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1 step multiplex probe kit/product/Solis BioDyne
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    1 step multiplex probe kit - by Bioz Stars, 2022-08
    94/100 stars
      Buy from Supplier

    Image Search Results


    Optimization, analytical sensitivity and clinical performance of rTEST COVID‐19/FLU qPCR kit. A. Heatmaps illustrate combinatorial testing of two IAV primer/probe sets at either high or low viral input (10 000 versus 10 copies per reaction). B. Heatmaps show combinatorial testing of IBV primer/probe sets at low viral input (10 copies per reaction). In panels A, B, the best performing primer/probe combinations (highlighted by green rectangles) were selected based on C t (darker colours denote higher sensitivity), fluorescent intensity (∆ R , lighter colours correspond to higher intensity) and the number of replicates that amplified. C. Analytical sensitivity of the multiplexed SARS‐CoV‐2 E and RdRP (both labelled with FAM), IAV PB1 and IBV PA (both labelled with YY), and RNase P assay. The dotted line at C t = 40 serves as a threshold after which amplification is considered invalid. D. Assessment of competitive interference of 1000 copies of IAV per reaction (500× LoD) on the analytical sensitivity of the SARS‐CoV‐2 E and RdRP assays multiplexed together. E. Assessment of competitive interference of 1000 copies of SARS‐CoV‐2 per reaction (500× LoD) on the analytical sensitivity of the IAV PB1 assay. F. Clinical performance of the rTEST COVID‐19/FLU qPCR kit. The dotted line and shaded area indicate samples that were not detected by a particular assay. C t , cycle threshold; E, envelope gene; IAV, influenza A; IBV, influenza B; PA, polymerase acidic protein; PB1, polymerase basic 1 protein; ND, not detected within 45 cycles; NTC, no template control; RdRP, RNA‐dependent RNA polymerase; Δ R , normalized fluorescent intensity.

    Journal: Microbial Biotechnology

    Article Title: Sequential development of several RT‐qPCR tests using LNA nucleotides and dual probe technology to differentiate SARS‐CoV‐2 from influenza A and B

    doi: 10.1111/1751-7915.14031

    Figure Lengend Snippet: Optimization, analytical sensitivity and clinical performance of rTEST COVID‐19/FLU qPCR kit. A. Heatmaps illustrate combinatorial testing of two IAV primer/probe sets at either high or low viral input (10 000 versus 10 copies per reaction). B. Heatmaps show combinatorial testing of IBV primer/probe sets at low viral input (10 copies per reaction). In panels A, B, the best performing primer/probe combinations (highlighted by green rectangles) were selected based on C t (darker colours denote higher sensitivity), fluorescent intensity (∆ R , lighter colours correspond to higher intensity) and the number of replicates that amplified. C. Analytical sensitivity of the multiplexed SARS‐CoV‐2 E and RdRP (both labelled with FAM), IAV PB1 and IBV PA (both labelled with YY), and RNase P assay. The dotted line at C t = 40 serves as a threshold after which amplification is considered invalid. D. Assessment of competitive interference of 1000 copies of IAV per reaction (500× LoD) on the analytical sensitivity of the SARS‐CoV‐2 E and RdRP assays multiplexed together. E. Assessment of competitive interference of 1000 copies of SARS‐CoV‐2 per reaction (500× LoD) on the analytical sensitivity of the IAV PB1 assay. F. Clinical performance of the rTEST COVID‐19/FLU qPCR kit. The dotted line and shaded area indicate samples that were not detected by a particular assay. C t , cycle threshold; E, envelope gene; IAV, influenza A; IBV, influenza B; PA, polymerase acidic protein; PB1, polymerase basic 1 protein; ND, not detected within 45 cycles; NTC, no template control; RdRP, RNA‐dependent RNA polymerase; Δ R , normalized fluorescent intensity.

    Article Snippet: RT‐qPCR reactions were optimized on a Mx3005P (Agilent Technologies), QuantStudio 5 (Agilent Technologies) and AriaMx (Agilent Technologies) real‐time PCR detection systems using SOLIScript® 1‐step CoV Kit (Cat. No. 08‐65‐00250; SOLIS BioDyne, Tartu, Estonia).

    Techniques: Real-time Polymerase Chain Reaction, Amplification

    Schematic illustrating SARS‐CoV‐2 genome and regions targeted by RT‐qPCR primers and probes. A. Schematic overview portrays the SARS‐CoV‐2 genome with RdRP and E gene regions magnified to show the locations of primers and probes of the original Charité protocol, vDetect (v1 and v2), and rTEST RT‐qPCR assays. F, forward primer; P, probe; R, reverse primer. The inset boxes (from left to right) illustrate a SARS‐CoV‐2 particle with labelled structural proteins and RNA, legend describing panel A and the primers and probes used in each test to detect RNase P subunit p30 (RPP30). B. Diagram compares the sequences of RdRP and E gene primers and probes for the original Charité protocol, vDetect (v.1) and vDetect v.2 and rTEST RT‐qPCR assays to the Wuhan reference sequence. The numbers written above the Wuhan reference sequence correspond to the start and end base positions of the sequence Reverse primer sequences are written in the reverse complement (rc). Magenta lines and letters represent mixed bases found in the primers and probes in the Charité protocol that were replaced with the correct bases in vDetect v1 (blue lines and letters). Red lines and letters signify LNA‐modified bases.

    Journal: Microbial Biotechnology

    Article Title: Sequential development of several RT‐qPCR tests using LNA nucleotides and dual probe technology to differentiate SARS‐CoV‐2 from influenza A and B

    doi: 10.1111/1751-7915.14031

    Figure Lengend Snippet: Schematic illustrating SARS‐CoV‐2 genome and regions targeted by RT‐qPCR primers and probes. A. Schematic overview portrays the SARS‐CoV‐2 genome with RdRP and E gene regions magnified to show the locations of primers and probes of the original Charité protocol, vDetect (v1 and v2), and rTEST RT‐qPCR assays. F, forward primer; P, probe; R, reverse primer. The inset boxes (from left to right) illustrate a SARS‐CoV‐2 particle with labelled structural proteins and RNA, legend describing panel A and the primers and probes used in each test to detect RNase P subunit p30 (RPP30). B. Diagram compares the sequences of RdRP and E gene primers and probes for the original Charité protocol, vDetect (v.1) and vDetect v.2 and rTEST RT‐qPCR assays to the Wuhan reference sequence. The numbers written above the Wuhan reference sequence correspond to the start and end base positions of the sequence Reverse primer sequences are written in the reverse complement (rc). Magenta lines and letters represent mixed bases found in the primers and probes in the Charité protocol that were replaced with the correct bases in vDetect v1 (blue lines and letters). Red lines and letters signify LNA‐modified bases.

    Article Snippet: RT‐qPCR reactions were optimized on a Mx3005P (Agilent Technologies), QuantStudio 5 (Agilent Technologies) and AriaMx (Agilent Technologies) real‐time PCR detection systems using SOLIScript® 1‐step CoV Kit (Cat. No. 08‐65‐00250; SOLIS BioDyne, Tartu, Estonia).

    Techniques: Quantitative RT-PCR, Sequencing, Modification

    Redesign and validation of Charité SARS‐CoV‐2 E and RdRP primer/probe sets. A. Performance of redesigned RdRP gene reverse primers with replaced mixed bases and optimized melting temperatures. B. Heatmap shows various E gene forward primers with and without LNA‐modified thymine residues (LNA‐T) and their relative performance amplifying SARS‐CoV‐2 template or samples contaminated with E gene synthetic positive control. C. Limit of detection of both E (left panel) and RdRP (right panel) gene assays. Dotted line at C t = 40 denotes the detection cut‐off. D. Clinical evaluation of both vDetect v.1 E (left panel) and RdRP (right panel) gene assays conducted in two independent laboratories. Dotted lines at ND and shaded areas show detection cut‐off and samples that were not detected for either the vDetect v.1 assay, index test assay, or both assays. C t , cycle threshold; E, envelope gene; ND, not detected within 45 cycles; NTC, no template control; R, reverse primer; RdRP, RNA‐dependent RNA polymerase.

    Journal: Microbial Biotechnology

    Article Title: Sequential development of several RT‐qPCR tests using LNA nucleotides and dual probe technology to differentiate SARS‐CoV‐2 from influenza A and B

    doi: 10.1111/1751-7915.14031

    Figure Lengend Snippet: Redesign and validation of Charité SARS‐CoV‐2 E and RdRP primer/probe sets. A. Performance of redesigned RdRP gene reverse primers with replaced mixed bases and optimized melting temperatures. B. Heatmap shows various E gene forward primers with and without LNA‐modified thymine residues (LNA‐T) and their relative performance amplifying SARS‐CoV‐2 template or samples contaminated with E gene synthetic positive control. C. Limit of detection of both E (left panel) and RdRP (right panel) gene assays. Dotted line at C t = 40 denotes the detection cut‐off. D. Clinical evaluation of both vDetect v.1 E (left panel) and RdRP (right panel) gene assays conducted in two independent laboratories. Dotted lines at ND and shaded areas show detection cut‐off and samples that were not detected for either the vDetect v.1 assay, index test assay, or both assays. C t , cycle threshold; E, envelope gene; ND, not detected within 45 cycles; NTC, no template control; R, reverse primer; RdRP, RNA‐dependent RNA polymerase.

    Article Snippet: RT‐qPCR reactions were optimized on a Mx3005P (Agilent Technologies), QuantStudio 5 (Agilent Technologies) and AriaMx (Agilent Technologies) real‐time PCR detection systems using SOLIScript® 1‐step CoV Kit (Cat. No. 08‐65‐00250; SOLIS BioDyne, Tartu, Estonia).

    Techniques: Modification, Positive Control

    Optimization of room temperature stable kit and dual probes for rTEST COVID‐19 qPCR kit. (A, B) Graphs depict the effects of decoy nucleic acids (tRNA or salmon sperm DNA) or pure oligonucleotides (pure) on the stability of lyophilized positive control left at room temperature over a one‐month period as assessed by amplification of RdRP (A) and E (B) genes. (C) Performance of either fresh or an rTEST COVID‐19 qPCR kit left a t room temperature for 1‐month on amplification of SARS‐CoV‐2 E and RdRP genes and human RNase P. Comparison of analytical sensitivity (D, F) and fluorescent intensity (E, G) between single probes versus dual probes for both RdRP (D, E) and E (F, G) genes. Dotted line at C t = 40 serves as a threshold after which amplification is considered invalid. * P

    Journal: Microbial Biotechnology

    Article Title: Sequential development of several RT‐qPCR tests using LNA nucleotides and dual probe technology to differentiate SARS‐CoV‐2 from influenza A and B

    doi: 10.1111/1751-7915.14031

    Figure Lengend Snippet: Optimization of room temperature stable kit and dual probes for rTEST COVID‐19 qPCR kit. (A, B) Graphs depict the effects of decoy nucleic acids (tRNA or salmon sperm DNA) or pure oligonucleotides (pure) on the stability of lyophilized positive control left at room temperature over a one‐month period as assessed by amplification of RdRP (A) and E (B) genes. (C) Performance of either fresh or an rTEST COVID‐19 qPCR kit left a t room temperature for 1‐month on amplification of SARS‐CoV‐2 E and RdRP genes and human RNase P. Comparison of analytical sensitivity (D, F) and fluorescent intensity (E, G) between single probes versus dual probes for both RdRP (D, E) and E (F, G) genes. Dotted line at C t = 40 serves as a threshold after which amplification is considered invalid. * P

    Article Snippet: RT‐qPCR reactions were optimized on a Mx3005P (Agilent Technologies), QuantStudio 5 (Agilent Technologies) and AriaMx (Agilent Technologies) real‐time PCR detection systems using SOLIScript® 1‐step CoV Kit (Cat. No. 08‐65‐00250; SOLIS BioDyne, Tartu, Estonia).

    Techniques: Real-time Polymerase Chain Reaction, Positive Control, Amplification

    Optimization, analytical sensitivity and clinical performance of a rapid, RNA extraction‐free, multiplexed RT‐qPCR assay. A. Analytical sensitivity of the triplexed E, RdRP and RNase P assay in the rTEST COVID‐19 qPCR Allplex kit. B. Clinical performance of the rTEST COVID‐19 qPCR Allplex kit. C. Optimization of gargle sample input for a rapid, RNA extraction‐free, triplexed rTEST. D. Comparison of four different thermal profiles using 8 μl of gargle input volume in rapid, direct RT‐qPCR. E. Analytical sensitivity of the triplexed E, RdRP and RNase P assay in the RNA extraction‐free rTEST COVID‐19 qPCR Rapid kit. F. Clinical performance of the rTEST COVID‐19 qPCR Rapid kit. The dotted line at C t = 40 (panels A and E) serves as a threshold after which amplification is considered invalid. The dotted lines and shaded areas (panels B and F) indicate samples that were not detected by either the evaluation test, index test or both tests. C t , cycle threshold; E, envelope gene; ND, not detected within 45 cycles; NTC, no template control; RdRP, RNA‐dependent RNA polymerase.

    Journal: Microbial Biotechnology

    Article Title: Sequential development of several RT‐qPCR tests using LNA nucleotides and dual probe technology to differentiate SARS‐CoV‐2 from influenza A and B

    doi: 10.1111/1751-7915.14031

    Figure Lengend Snippet: Optimization, analytical sensitivity and clinical performance of a rapid, RNA extraction‐free, multiplexed RT‐qPCR assay. A. Analytical sensitivity of the triplexed E, RdRP and RNase P assay in the rTEST COVID‐19 qPCR Allplex kit. B. Clinical performance of the rTEST COVID‐19 qPCR Allplex kit. C. Optimization of gargle sample input for a rapid, RNA extraction‐free, triplexed rTEST. D. Comparison of four different thermal profiles using 8 μl of gargle input volume in rapid, direct RT‐qPCR. E. Analytical sensitivity of the triplexed E, RdRP and RNase P assay in the RNA extraction‐free rTEST COVID‐19 qPCR Rapid kit. F. Clinical performance of the rTEST COVID‐19 qPCR Rapid kit. The dotted line at C t = 40 (panels A and E) serves as a threshold after which amplification is considered invalid. The dotted lines and shaded areas (panels B and F) indicate samples that were not detected by either the evaluation test, index test or both tests. C t , cycle threshold; E, envelope gene; ND, not detected within 45 cycles; NTC, no template control; RdRP, RNA‐dependent RNA polymerase.

    Article Snippet: RT‐qPCR reactions were optimized on a Mx3005P (Agilent Technologies), QuantStudio 5 (Agilent Technologies) and AriaMx (Agilent Technologies) real‐time PCR detection systems using SOLIScript® 1‐step CoV Kit (Cat. No. 08‐65‐00250; SOLIS BioDyne, Tartu, Estonia).

    Techniques: RNA Extraction, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Amplification

    CT value overview for 3600 clinical specimens tested with the RKI/ZBS1 SARS-CoV-2 protocol. CT values for the four PCR assays included in the RKI/ZBS1 SARS-CoV-2 protocol as determined for the first 3600 specimens. Red symbols show the SARS-CoV-2 assays (negatives not shown), blue squares the inhibition control KoMa and green circles the nucleic acid detection by c-myc amplification. The constant detection of the inhibition control KoMa can be seen, and the scattering of the nucleic acid content demonstrated by c-myc as well as the similar CT values for the E-Gene and the orf1ab assay

    Journal: Virology Journal

    Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

    doi: 10.1186/s12985-021-01559-3

    Figure Lengend Snippet: CT value overview for 3600 clinical specimens tested with the RKI/ZBS1 SARS-CoV-2 protocol. CT values for the four PCR assays included in the RKI/ZBS1 SARS-CoV-2 protocol as determined for the first 3600 specimens. Red symbols show the SARS-CoV-2 assays (negatives not shown), blue squares the inhibition control KoMa and green circles the nucleic acid detection by c-myc amplification. The constant detection of the inhibition control KoMa can be seen, and the scattering of the nucleic acid content demonstrated by c-myc as well as the similar CT values for the E-Gene and the orf1ab assay

    Article Snippet: Similarly, the duplexed c-myc assay seems to be more sensitive, with these reagents being positive in ten out of ten specimens with the SOLIScript® 1-step CoV kit, while the SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase amplifies c-myc only in two out of ten specimens from the same specimens.

    Techniques: Polymerase Chain Reaction, Inhibition, Amplification

    RKI/ZBS1 SARS-CoV-2 protocol performance on different PCR cyclers. To show that the presented protocol can be run on several cyclers, we used 10 clinical samples of different RNA load, negative as well as positive controls and set up one master mix that was distributed to the different cyclers as shown above. Different colors represent 10 different samples used for comparison. Mean values of duplicates are shown, also for SARS-CoV-2-positive controls (crosses); negative controls are not shown

    Journal: Virology Journal

    Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

    doi: 10.1186/s12985-021-01559-3

    Figure Lengend Snippet: RKI/ZBS1 SARS-CoV-2 protocol performance on different PCR cyclers. To show that the presented protocol can be run on several cyclers, we used 10 clinical samples of different RNA load, negative as well as positive controls and set up one master mix that was distributed to the different cyclers as shown above. Different colors represent 10 different samples used for comparison. Mean values of duplicates are shown, also for SARS-CoV-2-positive controls (crosses); negative controls are not shown

    Article Snippet: Similarly, the duplexed c-myc assay seems to be more sensitive, with these reagents being positive in ten out of ten specimens with the SOLIScript® 1-step CoV kit, while the SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase amplifies c-myc only in two out of ten specimens from the same specimens.

    Techniques: Polymerase Chain Reaction

    Correlation between the internal KoMa control and SARS-CoV-2 genome load in clinical specimens. For the 424 SARS-CoV-2-positive specimens the CT values were grouped as indicated and plotted against the CT value for the inhibition control KoMa. While in specimens that are highly positive for SARS-CoV-2 KoMa does not give a signal reliably, in specimens with genome loads close to the detection limit as well as in negative samples, KoMa is constantly amplified, showing the efficiency of the nucleic acid extraction and possible inhibitory effects of the sample matrix. Detectability increases significantly with E-Gene CT values higher than 25 (Mann Whitney p

    Journal: Virology Journal

    Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

    doi: 10.1186/s12985-021-01559-3

    Figure Lengend Snippet: Correlation between the internal KoMa control and SARS-CoV-2 genome load in clinical specimens. For the 424 SARS-CoV-2-positive specimens the CT values were grouped as indicated and plotted against the CT value for the inhibition control KoMa. While in specimens that are highly positive for SARS-CoV-2 KoMa does not give a signal reliably, in specimens with genome loads close to the detection limit as well as in negative samples, KoMa is constantly amplified, showing the efficiency of the nucleic acid extraction and possible inhibitory effects of the sample matrix. Detectability increases significantly with E-Gene CT values higher than 25 (Mann Whitney p

    Article Snippet: Similarly, the duplexed c-myc assay seems to be more sensitive, with these reagents being positive in ten out of ten specimens with the SOLIScript® 1-step CoV kit, while the SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase amplifies c-myc only in two out of ten specimens from the same specimens.

    Techniques: Inhibition, Amplification, MANN-WHITNEY

    Principle of the RKI/ZBS1 protocol for the real-time PCR detection of SARS-CoV-2. RKI/ZBS1 SARS-CoV-2 protocol

    Journal: Virology Journal

    Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

    doi: 10.1186/s12985-021-01559-3

    Figure Lengend Snippet: Principle of the RKI/ZBS1 protocol for the real-time PCR detection of SARS-CoV-2. RKI/ZBS1 SARS-CoV-2 protocol

    Article Snippet: Similarly, the duplexed c-myc assay seems to be more sensitive, with these reagents being positive in ten out of ten specimens with the SOLIScript® 1-step CoV kit, while the SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase amplifies c-myc only in two out of ten specimens from the same specimens.

    Techniques: Real-time Polymerase Chain Reaction

    RKI/ZBS1 SARS-CoV-2 protocol performance with different PCR master mixes. In total 7 PCR mixes were compared with the RKI/ZBS1 SARS-CoV-2 protocol on a Bio-Rad CFX96 cycler. 10 clinical specimens of different RNA load and negative as well as positive controls were set up. Means of duplicate CT values are plotted against the respective master mix. Positive controls (crosses) are shown only for the SARS-CoV-2 assays E-Gene and orf1ab; negative controls are not shown

    Journal: Virology Journal

    Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

    doi: 10.1186/s12985-021-01559-3

    Figure Lengend Snippet: RKI/ZBS1 SARS-CoV-2 protocol performance with different PCR master mixes. In total 7 PCR mixes were compared with the RKI/ZBS1 SARS-CoV-2 protocol on a Bio-Rad CFX96 cycler. 10 clinical specimens of different RNA load and negative as well as positive controls were set up. Means of duplicate CT values are plotted against the respective master mix. Positive controls (crosses) are shown only for the SARS-CoV-2 assays E-Gene and orf1ab; negative controls are not shown

    Article Snippet: Similarly, the duplexed c-myc assay seems to be more sensitive, with these reagents being positive in ten out of ten specimens with the SOLIScript® 1-step CoV kit, while the SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase amplifies c-myc only in two out of ten specimens from the same specimens.

    Techniques: Polymerase Chain Reaction

    Principle of the RKI/ZBS1 protocol for the real-time PCR detection of SARS-CoV-2. RKI/ZBS1 SARS-CoV-2 protocol

    Journal: Virology Journal

    Article Title: Resource-efficient internally controlled in-house real-time PCR detection of SARS-CoV-2

    doi: 10.1186/s12985-021-01559-3

    Figure Lengend Snippet: Principle of the RKI/ZBS1 protocol for the real-time PCR detection of SARS-CoV-2. RKI/ZBS1 SARS-CoV-2 protocol

    Article Snippet: To assess the performance of further commercially available one-step RT-PCR master mixes with the RKI/ZBS1 SARS-CoV-2 protocol, 10 clinical specimens with varying viral genome load were analyzed with the AgPath-ID master mix on the Bio-Rad CFX96 cycler and compared to the SuperScript™ III One-Step RT-PCR System with Platinum™ Taq DNA Polymerase (Invitrogen, Carlsbad, CA, USA), the lyophilized 1-step RT-PCR Polymerase Mix (TIB MOLBIOL, Berlin, Germany), the TaqPath™ 1-Step Multiplex Master Mix (No ROX) (Applied Biosystems), two kits from Solis BioDyne (SOLIScript 1-step Multiplex Probe Kit, SOLIScript® 1-step CoV Kit; Solis BioDyne, Tartu, Estonia) and the GoTaq® Probe 1-Step RT-qPCR System (Promega, Walldorf, Germany).

    Techniques: Real-time Polymerase Chain Reaction